The Brucella suis type IV secretion system assembles in the cell envelope of the heterologous host Agrobacterium tumefaciens and increases IncQ plasmid pLS1 recipient competence

被引:19
作者
Carle, A
Höppner, C
Aly, KA
Yuan, Q
den Dulk-Ras, A
Vergunst, A
O'Callaghan, D
Baron, C
机构
[1] McMaster Univ, Dept Biol, Hamilton, ON L8S 4K1, Canada
[2] Univ Munich, Bereich Mikrobiol, Dept Biol 1, D-80638 Munich, Germany
[3] Leiden Univ, Inst Biol, Clusius Lab, NL-2333 AL Leiden, Netherlands
[4] INSERM, Unite 431, UFR Med, F-30908 Nimes, France
关键词
D O I
10.1128/IAI.74.1.108-117.2006
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Pathogenic Brucella species replicate within mammalian cells, and their type IV secretion system is essential for intracellular survival and replication. The options for biochemical studies on the Brucella secretion system are limited due to the rigidity of the cells and biosafety concerns, which preclude large-scale cell culture and fractionation. To overcome these problems, we heterologously expressed the Brucella suis virB operon in the closely related alpha(2)-proteobacterium Agrobacterium tumefaciens and showed that the VirB proteins assembled into a complex. Eight of the twelve VirB proteins were detected in the membranes of the heterologous host with specific antisera. Cross-linking indicated protein-protein interactions similar to those in other type IV secretion systems, and the results of immunofluorescence analysis supported the formation of VirB protein complexes in the cell envelope. Production of a subset of the B. suis VirB proteins (VirB3-VirB12) in A. tumefaciens strongly increased its ability to receive IncQ plasmid pLS1 in conjugation experiments, and production of VirB1 further enhanced the conjugation efficiency. Plasmid recipient competence correlated with periplasmic leakage and the detergent sensitivity of A. tumefaciens, suggesting a weakening of the cell envelope. Heterologous expression thus permits biochemical characterization of B. suis type W secretion system assembly.
引用
收藏
页码:108 / 117
页数:10
相关论文
共 67 条
[11]   Definition of a bacterial type IV secretion pathway for a DNA substrate [J].
Cascales, E ;
Christie, PJ .
SCIENCE, 2004, 304 (5674) :1170-1173
[12]   The versatile bacterial type IV secretion systems [J].
Cascales, E ;
Christie, PJ .
NATURE REVIEWS MICROBIOLOGY, 2003, 1 (02) :137-149
[13]   Brucella coopts the small GTPase Sar1 for intracellular replication [J].
Celli, J ;
Salcedo, SP ;
Gorvel, JP .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (05) :1673-1678
[14]   Organelle robbery:: Brucella interactions with the endoplasmic reticulum [J].
Celli, J ;
Gorvel, JP .
CURRENT OPINION IN MICROBIOLOGY, 2004, 7 (01) :93-97
[15]   Type IV secretion:: the Agrobacterium VirB/D4 and related conjugation systems [J].
Christie, PJ .
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2004, 1694 (1-3) :219-234
[16]   Biogenesis, architecture, and function of bacterial type IV secretion systems [J].
Christie, PJ ;
Atmakuri, K ;
Krishnamoorthy, V ;
Jakubowski, S ;
Cascales, E .
ANNUAL REVIEW OF MICROBIOLOGY, 2005, 59 :451-485
[17]   IDENTIFICATION OF 7 SURFACE-EXPOSED BRUCELLA OUTER-MEMBRANE PROTEINS BY USE OF MONOCLONAL-ANTIBODIES - IMMUNOGOLD LABELING FOR ELECTRON-MICROSCOPY AND ENZYME-LINKED-IMMUNOSORBENT-ASSAY [J].
CLOECKAERT, A ;
DEWERGIFOSSE, P ;
DUBRAY, G ;
LIMET, JN .
INFECTION AND IMMUNITY, 1990, 58 (12) :3980-3987
[18]   Essential role of the VirB machinery in the maturation of the Brucella abortus-containing vacuole [J].
Comerci, DJ ;
Martínez-Lorenzo, MJ ;
Sieira, R ;
Gorvel, JP ;
Ugalde, RA .
CELLULAR MICROBIOLOGY, 2001, 3 (03) :159-168
[19]   The IncP plasmid-encoded cell envelope-associated DNA transfer complex increases cell permeability [J].
Daugelavicius, R ;
Bamford, JKH ;
Grahn, AM ;
Lanka, E ;
Bamford, DH .
JOURNAL OF BACTERIOLOGY, 1997, 179 (16) :5195-5202
[20]   Cellular bioterrorism:: how Brucella corrupts macrophage physiology to promote invasion and proliferation [J].
de Bagues, MPJ ;
Dudal, S ;
Dornand, J ;
Gross, A .
CLINICAL IMMUNOLOGY, 2005, 114 (03) :227-238